The Proteome of Circulating Large Extracellular Vesicles in Diabetes and Hypertension.

Hypertension and diabetes induce vascular injury through processes that are not fully understood. Changes in extracellular vesicle (EV) composition could provide novel insights. Here, we examined the protein composition of circulating EVs from hypertensive, diabetic and healthy mice. EVs were isolated from transgenic mice overexpressing human renin in the liver (TtRhRen, hypertensive), OVE26 type 1 diabetic mice and wild-type (WT) mice. Protein content was analyzed using liquid chromatography-mass spectrometry. We identified 544 independent proteins, of which 408 were found in all groups, 34 were exclusive to WT, 16 were exclusive to OVE26 and 5 were exclusive to TTRhRen mice. Amongst the differentially expressed proteins, haptoglobin (HPT) was upregulated and ankyrin-1 (ANK1) was downregulated in OVE26 and TtRhRen mice compared with WT controls. Conversely, TSP4 and Co3A1 were upregulated and SAA4 was downregulated exclusively in diabetic mice; and PPN was upregulated and SPTB1 and SPTA1 were downregulated in hypertensive mice, compared to WT mice. Ingenuity pathway analysis identified enrichment in proteins associated with SNARE signaling, the complement system and NAD homeostasis in EVs from diabetic mice. Conversely, in EVs from hypertensive mice, there was enrichment in semaphroin and Rho signaling. Further analysis of these changes may improve understanding of vascular injury in hypertension and diabetes.

Impact of acute moderate-intensity aerobic exercise on circulating extracellular vesicles in pregnant and non-pregnant women.

Exercise improves cardiovascular and metabolic health in pregnancy and may represent a non-pharmacological approach to improving pregnancy outcomes. Extracellular vesicles (EVs) are emerging biomarkers of endothelial dysfunction and offer the potential for evaluating vascular health non-invasively during pregnancy. The purpose of this study was to investigate changes in circulating EV levels after an acute bout of moderate-intensity aerobic exercise in healthy pregnant and non-pregnant women. We studied plasma samples from pregnant (N = 13, 13-28 weeks) and non-pregnant (N = 17) women. A pre-exercise blood sample was obtained followed by a 30 min bout of moderate-intensity treadmill-based exercise. Immediately following the exercise, a post-exercise blood draw was collected. Large EVs were isolated from plasma by differential centrifugation and characterized by Western blot and electron microscopy. We quantified circulating EVs by nanoscale flow cytometry. Endothelial EVs were identified as VE-Cadherin+, platelet EVs as CD41+, and leukocyte EVs as CD45+ events. Acute exercise was associated with a significant reduction in levels of circulating endothelial EVs in the non-pregnant group (p = 0.0232) but not in the pregnant group (p = 0.2734). A greater proportion of non-pregnant women (13/17, 76.47%) exhibited a reduction in endothelial EVs compared with their pregnant counterparts (4/13, 30.76%, p < 0.05). We also observed a positive association between measures of fitness (average speed) and baseline levels of platelet (r = 0.5816, p = 0.0159) and total EVs (r = 0.5325, p = 0.0296) in the non-pregnant group but not in pregnant individuals. Collectively, our study highlights that after a matched acute exercise, changes to circulating EV levels differ depending on pregnancy status.

Extracellular vesicles in gestational diabetes mellitus: A scoping review.

Gestational diabetes mellitus (GDM) is one of the most common complications of pregnancy worldwide. Despite extensive study, the molecular mechanisms leading to GDM and associated perinatal complications are not well understood. The condition is also associated with an increased risk of future cardiometabolic disease in both mothers and their offspring. Thus, there is a pressing need for the development of effective screening tools and to identify novel molecular mechanisms responsible for the short and long-term risks associated with GDM. In this regard, extracellular vesicles (EVs) offer promise as novel biomarkers of GDM-mediated changes to both mother and fetus. The purpose of this scoping review is to provide an overview of studies examining EVs in the context of GDM. EMBASE and Ovid Medline were searched for articles published from inception to December 2020. We update current knowledge in this area and identify key knowledge gaps with recommendations for future research.

Circulating extracellular vesicles during pregnancy in women with type 1 diabetes: a secondary analysis of the CONCEPTT trial.

BACKGROUND: Extracellular vesicles are membrane vesicles that are released into the extracellular environment and accumulate in the circulation in vascular disease. We aimed to quantify circulating extracellular vesicles in pregnant women with type 1 diabetes and to examine associations between extracellular vesicle levels, continuous glucose measures, and pregnancy outcomes. METHODS: We used plasma samples from the Continuous Glucose Monitoring in Women with Type 1 Diabetes in Pregnancy Trial study and quantified circulating extracellular vesicles by flow cytometry (n = 163). Relationships with clinical variables were assessed by repeated measures correlation. Logistic regression was used to assess associations between elevated extracellular vesicle levels and pregnancy outcomes. RESULTS: Platelet extracellular vesicle levels were inversely associated with glucose time above range and glycaemic variability measures (P < 0.05). A weak positive association was observed between endothelial extracellular vesicles and mean amplitude of glycemic excursion (P < 0.05). In a univariate logistic regression model, high baseline endothelial extracellular vesicles was associated with increased risk of neonatal intensive care unit (NICU) admission (OR: 2.06, 1.03-4.10), and respiratory distress requiring ventilation (OR: 4.98, 1.04-23.92). After adjusting for HbA1c and blood pressure the relationship for NICU admission persisted and an association with hyperbilirubinemia was seen (OR: 2.56, 1.10-5.94). Elevated platelet extracellular vesicles were associated with an increased risk of NICU admission (OR: 2.18, 1.04-4.57), and hyperbilirubinemia (OR: 2.61, 1.11-6.12) after adjusting for HbA1c and blood pressure. CONCLUSIONS: High levels of extracellular vesicles in early pregnancy were associated with adverse neonatal outcomes. Assessment of extracellular vesicles may represent a novel approach to personalized care in type 1 diabetes pregnancy.

How Changes in Anti-SD Sequences Would Affect SD Sequences in Escherichia coli and Bacillus subtilis.

The 3' end of the small ribosomal RNAs (ssu rRNA) in bacteria is directly involved in the selection and binding of mRNA transcripts during translation initiation via well-documented interactions between a Shine-Dalgarno (SD) sequence located upstream of the initiation codon and an anti-SD (aSD) sequence at the 3' end of the ssu rRNA. Consequently, the 3' end of ssu rRNA (3'TAIL) is strongly conserved among bacterial species because a change in the region may impact the translation of many protein-coding genes. Escherichia coli and Bacillus subtilis differ in their 3' ends of ssu rRNA, being GAUCACCUCCUUA3' in E. coli and GAUCACCUCCUUUCU3' or GAUCACCUCCUUUCUA3' in B. subtilis Such differences in 3'TAIL lead to species-specific SDs (designated SDEc for E. coli and SDBs for B. subtilis) that can form strong and well-positioned SD/aSD pairing in one species but not in the other. Selection mediated by the species-specific 3'TAIL is expected to favor SDBs against SDEc in B. subtilis, but favor SDEc against SDBs in E. coli Among well-positioned SDs, SDEc is used more in E. coli than in B. subtilis, and SDBs more in B. subtilis than in E. coli Highly expressed genes and genes of high translation efficiency tend to have longer SDs than lowly expressed genes and genes with low translation efficiency in both species, but more so in B. subtilis than in E. coli Both species overuse SDs matching the bolded part of the 3'TAIL shown above. The 3'TAIL difference contributes to the host specificity of phages.